Lambda DNA prep.
robin_beech at maclan.mcgill.ca
Tue Feb 20 09:15:12 EST 1996
In article <Schenker.26.04FE43B0 at MPIMG-Berlin-Dahlem.MPG.DE>,
Schenker at MPIMG-Berlin-Dahlem.MPG.DE (Martin Schenker) wrote:
> Hi to everybody!!
> I'm working with a DASH II-library and struggle with the normal technique of
> DNA prepping, like DNase/RNase, PEG, Prot. K, SDS, EtoH.
> Is there a fast and reliable method to get good DNA (quality and
> of these little ********??
> I have to check many of them, so if it is fast, it would be preferred.
> Thanks for any suggestion,
I used to use a fairly simple prep which I haven't seen anywhere else
1.5 ml overnight liquid lysate into an eppendorf and microfuge
Transfer 0.8 ml to a new tube and make 1µg per ml DNAse
Incubate at room temperature for 15 min
Add 0.2 ml of TES (Tris 50 mM pH 8.0; EDTA 10 mM; SDS 5%)
Incubate at 70 deg C 15 min
Cool on ice and add 0.15 ml 8M K acetate
Microfuge then extract the supernatent with phenol-chloroform
Add isopropanol and recover the DNA precipitate
Hope it helps
Institute of Parasitology
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